MicroRNAs (miRNA) are short, noncoding RNAs with pervasive roles throughout gene expression in cellular processes such as differentiation and disease states such as cancer. Uncovering the roles of these molecules in development and tumorigenesis are key steps to the discovery of robust, new biomarkers and potential disease cures.
Existing methods to analyze miRNA expression consist of single-target PCR based methods and highly multiplexed array methods. PCR is very sensitive but can only analyze a single target. Thus, detection of miRNA panels requires large numbers of parallel reactions, greatly increasing cost, sample consumption, and complexity. Existing array based methods can analyze hundreds of miRNA in a single reaction but have high cost per sample, low sample throughput, and limited sensitivity. Successful clinical validation and translation of these promising miRNA panels will require that 10-100 miRNA be accurately quantified in a cost-effective, high-throughput, and robust manner. No existing technology can bridge this gap between the highly multiplexed but expensive array-based methods and the sensitive but singleplex qPCR methods. Currently, no method is able to achieve the high sensitivity, multiplex detection necessary for low cost miRNA profiling in rare clinical samples.
Thus, there exists a need for a low cost, PCR-free, multiplexed assay for miRNA expression analysis that performs highly quantitative, 10-100 plex miRNA profiling in a single reaction in less time than currently existing methods.